I am synthesizing Albumin nano particles for drug delivery. In every protocol, glutaraldehyde was added to the solution for crosslinking. Why is the crosslinking so important for albumin nano particles? And how does glutaraldehyde promotes it?
Glutaraldehyde is a homobifunctional crosslinker that is very reactive, so frequently if you allow too much crosslinking then you lose enzyme activity, so you need to optimize to have crosslinking for stabilization, but not complete loss of activity.
If you have a look at the attached literature ( Results and discussion first paragraph), it can be helpful to find the answer.
Hi Abdul,
glutaraldehyde is a very reactive compound and reacts unselectively with e.g. amino groups of proteins forming amide bonds. The protein is then usuallynot functional any more since reactive compartments are "glued" together. It can even yield to crossling of several protein chains to form gel particles (that's what you want).
I attached two links regarding the chemistry behind GA crosslinking.
Marcus
http://www.sciencedirect.com/science/article/pii/0003986168905547
http://www.sciencedirect.com/science/article/pii/0019279169901785
Glutaraldehyde is a homobifunctional crosslinker that is very reactive, so frequently if you allow too much crosslinking then you lose enzyme activity, so you need to optimize to have crosslinking for stabilization, but not complete loss of activity.
Another thing is that you should store and open the glutaraldehyde vial/bottle only under Argon (use a big funnel with Argon flushing through for that, you can place the bottle in the Argon stream). Since it is a very reactive compound, it also oxidizes with time to glutaric acid (consuming oxygen from air), which is then not reactive any more. If I want to be sure, I open a fresh new vial rather than using an old one from somewhere in the back of the fridge... :)
@Marcus,
So, can we transfer glutaraldehyde into vials in argon gas environment, as different batches and store them for future purpose? Because, not all the laboratories are equipped with argon gas environment chamber
If you keep them tightly closed until use in the fridge, this will be no problem. If you open it once and close it under air again, that won't be a big issue. If you do it more often, it will oxidize to complete conversion with time.
Small single-use aliquots are best then.
Hi,
I'd like to know your comments toward EDC as a glutaraldehyde alternative, since I have read an article* using EDC. It surmised 1)the EDC is less toxic for it's a zero space cross-linker 2) therefore, easily separated by centrifugation 3) more efficient: 3 hr instead of overnight.
EDC seems to be a good crosslinking agent, but why glutaraldehyde is still the majority?
*A simple improved desolvation method for the rapid preparation of albumin nanoparticles
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